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Application Note: Combining Small-Scale Purification and Analysis of Monoclonal Antibodies on One Instrument

27 November 2012

This application note presents a method for the small-scale purification of a monoclonal antibody from a cell lysate and the subsequent analysis on a single instrument, the Agilent 1260 Infinity Bio-Inert Quaternary LC System. Purification and polishing steps using preparative columns for Protein A purification and size exclusion chromatography were combined with high-volume injection using the Agilent 1260 Infinity Bio-Inert Manual Injector. Analysis of the purified mAb is demonstrated using the Agilent Protein A Monolith for quantification and identification. In addition, charge variant analysis was carried out by a four-component ionic strength gradient using ion exchange chromatography, calculated with Agilent Buffer Advisor software.